The overall objective of this preclinical research project is to assess the efficacy of benzyl isothiocyanate (BITC), a constituent of many edible cruciferous vegetables for prevention of pancreatic cancer using in vitro cell culture and in vivo animal models. The rationale for these studies stems from recent epidemiological data as well as preliminary results obtained in our laboratory. Several epidemiological studies have supported that frequent intake of cruciferous vegetables is linked to a decreased risk in various type of human cancer including pancreatic carcinoma. Our preliminary studies in vitro led us to a novel hypothesis that BITC induced death receptor as well as phosphorylation of antiapoptotic protein Bcl-xL might be predominant factors to destroy pancreatic cancer cells and prevent its progression. BITC might behave as an agent that is capable of intervening simultaneously at several targets in the carcinogenic process. This type of compound is thought to have selective advantage over other single-target compounds. Specific Aim 1 will systemically determine the role of intrinsic (phosphorylation of Bcl-xL and involvement of mitochondria) and extrinsic (death receptor mediated activation of caspase-8) pathways in BITC mediated apoptosis induction in pancreatic cancer cells. In specific Aim 2, the effect of oral administration of BITC on the progression of Pancreatic Intraepithelial Neoplasia (Pan IN) in a conditional KrasG12D mouse model will be investigated. It is believed that pancreatic ductal adenocarcinoma gradually arises from precursor lesion Pan IN. In that respect conditional KrasG12D mouse model is a valuable tool to investigate Pan IN biology. Biochemical assays will also be deployed to assess whether in vitro studies such as death receptor up regulation or Bcl-xL phosphorylation can be translated in vivo if any delay or prevention of the progression of Pan IN occurs in BITC treated animals. In summary, the studies proposed in this research program never explored before will I) define the mechanism by which BITC inhibits growth of human pancreatic cancer cells that may be helpful to identify mechanism- based biomarkers for future clinical trials and II) assess in vivo efficacy of BITC against pancreatic cancer using established animal model, which is a prerequisite for ensuing any clinical trial with this compound against pancreatic cancer. PUBLIC HEALTH RELEVANCE: Project Narrative At present, pancreatic cancer ranks as the fourth leading cause of cancer mortality in the United States (U.S.). The long-term goal of this preclinical research project is to assess the efficacy of benzyl isothiocyanate (BITC), a constituent of many edible cruciferous vegetables for prevention of pancreatic cancer using in vitro as well as animal models.